GSM7178915: Organoids_sh4-1; Homo sapiens; RNA-Seq
The total RNA was processed using mRNA enrichment, which involved enriching for mRNA with polyA tails using magnetic beads with OligodT. The obtained RNA was fragmented using a fragmentation buffer, and reverse transcription was performed with random N6 primers to synthesize cDNA double strands. The synthesized double-stranded DNA was end-repaired and 5'-phosphorylated, and the 3'-end was extended with an "A" tail to form a sticky end. A bubble-shaped adapter with a protruding "T" at the 3' end was then ligated to the DNA fragments, which were PCR amplified using specific primers. The PCR products were denatured to form single-stranded DNA, which was circularized using a bridge primer to obtain a single-stranded circular DNA library.